Quick development of disease-model microminiature pig by RNA-guided gene drive

2018 Fiscal Year Final Research Report

• Project/Area Number: 17K19326
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Veterinary medical science, Animal science, and related fields
• Research Institution: Kyushu University
• Principal Investigator: ONO ETSURO 九州大学, 医学研究院, 教授 (00160903)
• Co-Investigator(Kenkyū-buntansha): 大竹 正剛 静岡県畜産技術研究所, 中小家畜研究センター 養豚・養鶏, 上席研究員 (90605677)
• Research Collaborator: TOMIOKA yukiko ; ENYA satoko ; KANGAWA akihisa
• Project Period (FY): 2017-06-30 – 2019-03-31
• Keywords: gene drive / ゲノム編集 / Msh2遺伝子 / リンチ症候群 / 疾患モデルミニブタ / マイクロミニピッグ

Outline of Final Research Achievements

Plasmid for RNA-guided gene drive of the Msh2 gene was constructed. The plasmid, sgRNA and Cas9 protein were transfected into embryonic fibroblasts (PEF) of microminiature pig (MMP). Three different cell lines destroyed the Msh2 gene by genome editing were established. RNA-guided gene drive of the Msh2 gene using fertilized embryos of MMP was performed twice. The plasmid, sgRNA and Cas9 protein were co-microinjected into the pronuclei of fertilized MMP embryos. These embryos were subsequently transplanted into the oviducts of pseudopregnant foster recipient pigs. In one of the foster recipient pigs, the pregnancy was confirmed. It is, therefore, that the birth of genome-edited MMP are expected.

Free Research Field

実験動物学

Academic Significance and Societal Importance of the Research Achievements

Gene driveによる遺伝子改変マイクロミニピッグ（MMP）の作製方法の確立は、迅速かつ効率的な遺伝子改変MMPの提供を可能とし、がん・免疫・神経・発生等の高次生命システム研究を加速度的に推進させ、創薬産業へも成果を還元できる。
リンチ症候群は大腸がんや子宮内膜、卵巣、胃、小腸、肝胆道系、腎盂・尿管がんなどの発症リスクが高まる疾患で、全大腸がんの2-5％程度がリンチ症候群と考えられ、最も頻度が高い遺伝性腫瘍の一つである。従って、リンチ症候群の中型動物モデルとして、Msh2遺伝子を欠損させたリンチ症候群MMPを開発することは、本疾患に対する新規予防治療法の開発や外科手術法の開発に貢献する。